Results
This is our results1. The expression of α-tubulin、β-tubulin、n-luciferase、c-luciferase
1.1 vector construction
We got gene fragments of α-tubulin、β-tubulin、n-luciferase、c-luciferase via PCR and verified them by electrophoresis(fig.) The theoretic gene size of α-tubulin is 1356bp, β-tubulin is 1335bp, n-luciferase is 1248bp, c-luciferase is 459bp, which fit our experimental results.
We ligated our gene fragments to E.coli expression plasmid pET30a(+) and the sequencing result showed that we successfully constructed the α-tubulin, β-tubulin, n-luciferase, c-luciferase expression vectors.
We transformed the expression vectors to E.coli expression strain TranB(DE3). After culturing and inducing with IPTG, we lysed the bacteria and used SDS-PAGE/ western-blot to test the protein from the bacteria supernatant, pellet and the renatured inclusion body.
Apart from this, we also transformed plasmids to Rosetta(DE3) which can express rare codons and improve the expression level of eukaryotic protein.
Based on the results, α-tubulin and β-tubulin were successfully expressed in cell.
We collaborated with Fujian Agriculture and Forest University and asked them to test the interaction between α and β-tubulin. Thus verified the activity of tubulin monomers.
这里放发福的结果